中文摘要：

构建 pPIC9K-VEGF165分泌型载体，线性化后电击转化至 GS115（his4）中，经最小葡萄糖培养基（MD 平板）筛选出阳性表达菌株并进行聚合酶链式反应（PCR）验证．菌株发酵上清液经 Sephadex G-25，Hep-arin Sepharose FF 和 Sephacryl S-100层析介质分离纯化，目的蛋白纯度达到95％，相对分子质量约为24ku．结果表明：重组人 VEGF165蛋白能诱导人脐静脉内皮细胞（HUVEC）增殖，提高 HUVEC 细胞的活性；重组人VEGF165蛋白免疫小鼠，制备多克隆抗体，间接酶联免疫吸附测定法（ELISA）检测抗体效价达1：51200．

英文摘要：

The recombinant secretory vector of pPIC9K-VEGF165 was constructed,linearized and then transformed into Pichia pastoris strain GS115 （his4）by electroporation.The positive transformants were seperated by minimal dextrose medium （MD plate）plate screening and confirmed by polymerase chain reaction （PCR）.Subsequently,the rhVEGF165 protein was obtained by purification from the supernatant with Sephadex G-25,Heparin Sepharose FF and Sephacryl S-100 gel filtration chromatograph with relative molecular mass of 24 ku and purity of 95%.Furthermore,the recombinant protein was found to have high potency of inducing HUVEC cell proliferation and improving human umbilical vein endo-thelial cell （HUVEC）cell activity.The polyclonal antibody generated from rhVEGF165 immunized mouse exerted the ti-ter of 1∶51 200 by indirect ELISA analysis.